Kansas State University researchers have discovered how weeds develop resistance to the popular herbicide glyphosate, a finding that could have broad future implications in agriculture and many other industries.

Their work is detailed in an article that appears in the Proceedings of the National Academy of Sciences, known as PNAS and considered to be one of the most-cited journals for scientific research in the world. According to its website, PNAS receives more
than 21 million hits per month.

"Herbicide resistance in weeds has been a huge problem, not only in Kansas and the U.S. but many parts of the world," said Mithila Jugulam, a K-State weed scientist and co-author of the PNAS article.

"What we found that was new was how these weeds have evolved resistance to glyphosate in such a short time. If you look at the evolution of glyphosate resistance in Palmer amaranth, based on our research, it appears to have occurred very rapidly."

Funding for this research was provided in part by grants from the Kansas Wheat Commission; the Kansas Crop Improvement Association; a National Science Foundation grant received through the Wheat Genetics Resource Center; the K-State Department of
Agronomy (College of Agriculture); and USDA's Agricultural Research Service. Kansas State University worked in collaboration with researchers at Clemson University, the USDA Agricultural Research Service (Mississippi) and Michigan State University.

Palmer amaranth and common waterhemp are the two troublesome pigweeds in Kansas agricultural fields, as well as other parts of the United States. Glyphosate -- the key ingredient in the popular Roundup brand -- is the herbicide that is widely used for
controlling many weeds. But Jugulam notes that glyphosate resistance is becoming more prevalent in many states.

"We found that glyphosate-resistant Palmer amaranth plants carry the glyphosate target gene in hundreds of copies," Jugulam said. "Therefore, even if you applied an amount much higher than the recommended dose of glyphosate, the plants would not be killed."

Bikram Gill, director of Kansas State University's Wheat Genetics Resource Center who has worked in plant genetics for nearly 50 years, said the researchers knew pretty quickly that the genetic makeup of resistant weeds was different.

"Normally, the genetic material in all organisms -- including humans -- is found in long, linear DNA molecules, called chromosomes," said Gill, another co-author of the study. "But when (K-State researchers) Dal-Hoe Koo and Bernd Friebe, the chromosome
experts on the team, looked at these glyphosate-resistant weeds, the glyphosate target gene, along with other genes actually escaped from the chromosomes and formed a separate, self-replicating circular DNA structure."

Scientists refer to this structure as extra-chromosomal circular DNA (eccDNA). Each eccDNA has one copy of the gene that produces an enzyme that is the target for glyphosate.

"Because of the presence of hundreds of eccDNAs in each cell, the amount of the enzyme is also abundant," Gill said. "Therefore, the plant is not affected by glyphosate application and the weed is resistant to the herbicide."

Gill said the indications are that once a weed has acquired eccDNA, the resistance may evolve as quickly as in one generation.

"We think that the resistance via eccDNA is transitory: It can be passed to the weed's offspring and other related weed species," he said. "We have somehow caught it in between becoming permanently resistant. Eventually, we think that these eccDNAs can be
incorporated into the linear chromosome. If that happens, then they will become resistant forever."

The same K-State group recently published research on common waterhemp in the scientific journal, Plant Physiology, reporting that "a portion of the linear chromosome containing the target gene broke to form a ring chromosome carrying several copies of the
glyphosate target gene," according to Jugulam.
Armed with their new knowledge, the researchers can begin work on developing strategies to negate resistance in weeds.

"It's been known that these circular DNA/chromosomal structures can be unstable," Jugulam said. "What we want to explore is, for example, if we do not apply glyphosate repeatedly or reduce the selection by glyphosate, can we make these ring-structured
chromosomes unstable and once again make these plants susceptible to glyphosate."

The research team notes that farmers should incorporate best management strategies -- such as rotating herbicides and crops -- to reduce weed pressure: "This may allow evolving resistance to dissipate as we know that these eccDNAs and ring chromosomes
are unstable and can be lost in the absence of herbicide selection pressure," Jugulam said.

"Glyphosate has a lot of good characteristics as an herbicide molecule," she added. "The recommendations that K-State and many others are promoting is 'do not abuse glyphosate.' Use the recommended integrated weed management strategies so that we do
not lose the option of using glyphosate for the sustainability of our agriculture."

Gill's distinguished career of improving the world's wheat varieties has spanned nearly a half century.

So it's a big deal when he sees a discovery that he feels truly changes the landscape of growing farm crops.

Gill and fellow scientists Dal-Hoe Koo and Bernd Friebe are among the co-authors of a study released on March 12 in which researchers found the mechanism that makes weeds resistant to glyphosate, the herbicide most commonly used in agriculture.

Gill's group worked alongside Mithila Jugulam, a weed scientist in the university's department of agronomy, in the discovery of a novel structure in the chromosome of resistant weeds.

"I have spent all of my life working on wheat and bread wheat, where we have founded many technologies," Gill said. "When we started working with Mithila on weeds, we said 'my goodness, all of the genetics we learned has gone out the window.' We have
discovered new genetic elements in weeds unknown to science, and It has been an absolute blast and incredible journey."

Gill added: "It just tells you that if you can work together and bring different disciplines together, you can do some wonderful science."